We propose to purchase a Coherent AntiStokes Raman Scattering Microscopy system. This instrument offers novel molecule specific label free imaging of biological samples and tissue. In particular this system will be compatible with intravital microscopy approaches and will be tunable such that it can detect multiple molecules in the same sample. A wide range of investigators involved in NIH clinical and basic research propose to use the system to enhance and develop their interests. These individual projects exploit the full range of benefits that CARS brings to imaging. These include label free non invasive imaging, compatibility with live cell and intravital microscopy. The NIR laser source used in this system is particularly suitable for tissue imaging. Depth resolved imaging is inherent in the CARS technique. This allows mapping of three dimensional tissue or sample volumes with submicron resolution. The CARS microscope has been specified by a diverse range of investigators working on a range of disease related problems. The strength of CARS in imaging lipid deposits and lipid dynamics has been fully exploited by this group. In addition we also propose its use in imaging calcifications such as in the vasculature. The CARS system adds new breadth to research capabilities in the basic sciences and medicine. Its location on a medical research campus ensures its effective and imaginative use. PUBLIC HEALTH RELEVANCE: CARS microscopy is a new method for imaging the three dimensional distribution of target molecules occurring in cells and tissue without the needing to apply an external marker to those molecules. Imaging of live cells and tissues is possible which in turn leads to new understanding of the role of these molecules in dynamic physiological processes and in disease.